A simple alkali lysis method for Plasmodium falciparum DNA extraction from filter paper blood samples

• Published in: Molecular and biochemical parasitology 2023-06, Vol.254, p.111557-111557, Article 111557
• Main Authors: Xiang, Zheng, Li, Danlin, Wang, Siqi, Shen, Ting, He, Wen, Li, Mier, Zeng, Weilin, Chen, Xi, Wu, Yanrui, Cui, Liwang, Yang, Zhaoqing
• Format: Article
• Language: English
• Subjects: Alkali lysis; DNA extraction; DNA, Protozoan - genetics; Humans; Malaria, Falciparum - parasitology; Parasitemia; Plasmodium falciparum; Plasmodium falciparum - genetics; Sensitivity and Specificity; Sodium Hydroxide
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2023.111557

A fast, simple, easy, efficient, and inexpensive method for DNA extraction from malaria parasites collected on filter paper would be very useful for molecular surveillance. The quality and quantity of DNA are critical to molecular diagnosis and analysis. Here, we developed a simple alkali lysis method for DNA extraction from blood samples on filter paper. The results showed that 10–50 mM NaOH and deionized water all effectively isolated parasite DNA at higher parasitemia, as witnessed by successful PCR amplification, while at a parasitemia of 0.01%, the 10 mM NaOH lysis condition generated the best results. Furthermore, DNA extracted by this method was successfully used to amplify a fragment of > 2000 bp. This method successfully extracted DNA from 1 µl of blood at a parasitemia as low as 0.0001% (equivalent to 5 parasites /µl). The DNA isolated by the 10 mM NaOH lysis method was stable to yield PCR products after storage at 4 °C or − 20 °C for 12 months. These results indicate that this alkali lysis method is simple, effective, sensitive, and inexpensive for isolating stable Plasmodium DNA from dried blood spots on filter paper. •Alkali lysis method was efficient for malaria molecular research from filter paper blood samples.•The most suitable NaOH concentration was 10 mM when extraction Plasmodium DNA from low parasitemia filter paper.•The present method enables detection of low parasitemia (0.0001% parasitemia, equivalent to 5 parasites /µl) sample.•The DNA isolated by alkali lysis method is more stable when stored at − 20 °C.